Setting up incubator for polyp bailout experiment, part II

date created 20210611; date last revised 20210614 removed moon light from hydra 52ai light type list

this post is the sequel to an earlier post about incubator setup planning

the previous post (part I) was for note taking on incubator dimensions and potential tank configuration

this post describes the ‘final’ configuration. This includes light, temperature, and tank/carboy configuration

All containers with water (carboys, bailout tanks, 6well plates) are kept in plastic bins to prevent flooding. All plastic bins are white so bin color (and potential changes in light reflected) is constant among treatments. All electronics are on the top shelf to prevent water damage. Pictures below

temperature - set to ~28.5C (results in ~26.5C water temp inside tanks which is similar to the water tables)

Light (2 hydra AI 52 lights)

Parameters-Ramp-Schedule

note: want to keep light constant for settlement experiments but the 10min ramp was the shortest option

  • Morning Ramp: 6am-6:10am
  • Peak: 6:10am-6pm
  • Evening Ramp: 6pm-6:10pm

Light-Settings

Lights were set so that the light levels near the 6 well settlement plates and the bailout containers is 30-40 umol/m2/s. Info on water table light conditions here.

Type of light Percent - top light Percent - bottom light
UV 3% 2%
Violet 3% 2%
Royal 3% 2%
Blue 3% 2%
Green 3% 2%
Deep Red 3% 2%
Cool Light 3% 2%

Carboys

Two 2-gallon carboys are on the bottom shelf in a plastic bin. One carboy has mixing tank seawater (~32.5psu, not from the recirculating tanks so elemental composition is known). The high salinity carboy has mixing tank seawater with extra Fritz RPM salt so the salinity is adjusted to ~78psu. Seawater from both was filtered over a 0.22um filter.

Tanks for Bailout

  • 2L clear, circular plastic containers (height: 10in, diameter: 4.25in)

Mesh bailout chambers

  • Plastic salad dressing to-go cups with 150um mesh holes in the bottom and 3 around the side. After the mesh was hot glued onto the plastic the chambers were soaked in water for 30min so leachate is completed before they come in contact with corals.

Lessons learned during bailout chamber construction

  • small lengths (~1in) of pvc and plastic tubing (~1in diameter) sank. We had some success floating these with egg crate (taped some of the small squares and the perimeter of the egg crate to improve buoyancy but the egg crate took up too much space which would allow for fewer bailout chambers/tank). We opted not to float pvc or plastic tubing with styrofoam to prevent contamination. Wrapping aquarium tubing around the plastic or pvc as “floaties” did not work either (both ends of the plastic tubing were taped over to prevent flooding).

we will rotate tank and 6well plate configuration everyday to account for variation in light intensity

photos

top of incubator with 6well settlement plates

bottom of incubator with carboys and bailout tanks

close up of 2L bailout tank with mesh chambers

Written on June 11, 2021